REL 2023-04-25 2023-06-23 2023-06-23 0000-0003-3693-2531 Robert A Dick Cornell University 360 Biotechnology Building, 526 Campus Road Ithaca New York United States 14853 0000-0003-3693-2531 Robert A Dick Cornell University 360 Biotechnology Building, 526 Campus Road Ithaca New York United States 14853 Highland CMH Dick RAD National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID) R01AI147890 United States 2.0 10.6019/EMPIAR-11571 EMDB molecule EMD-29773 Highland CM Tan A Ricaña CL Briggs JAG Dick RA Hurley JHH Proceedings of the National Academy of Sciences of the United States of America Proc Natl Acad Sci U S A United States 18 120 2023 English 10.1073/pnas.2220545120 37094124

ABSTRACT: The HIV-1 capsid houses the viral genome and interacts extensively with host cell proteins throughout the viral life cycle. It is composed of capsid protein (CA), which assembles into a conical fullerene lattice composed of roughly 200 CA hexamers and 12 CA pentamers. Previous structural analyses of individual CA hexamers and pentamers have provided valuable insight into capsid structure and function, but detailed structural information about these assemblies in the broader context of the capsid lattice is lacking. In this study, we combined cryoelectron tomography and single particle analysis (SPA) cryoelectron microscopy to determine structures of continuous regions of the capsid lattice containing both hexamers and pentamers. We also developed a method of liposome scaffold-based in vitro lattice assembly (“lattice templating”) that enabled us to directly study the lattice under a wider range of conditions than has previously been possible. Using this approach, we identified a critical role for inositol hexakisphosphate in pentamer formation and determined the structure of the CA lattice bound to the capsid-targeting antiretroviral drug GS-6207 (lenacapavir). Our work reveals key structural details of the mature HIV-1 CA lattice and establishes the combination of lattice templating and SPA as a robust strategy for studying retroviral capsid structure and capsid interactions with host proteins and antiviral compounds.

Unaligned multi-frame micrographs of HIV-1 CA lattice templated on 30-nm liposomes in the presence of assembly cofactor IP6 at pH 6.2 /data/temp6_tif-frames_fixed micrographs - multiframe TIFF TIFF 696 50 UNSIGNED BYTE 11520 0.655 8184 0.655

'Raw,' unaligned multi-frame movies collected using a Talos Arctica microscope equipped with a Gatan K3 detector/BioQuantum energy filter operating in super-resolution mode. Voltage: 200 kV Physical pixel size: 1.31Å/pixel Total dose: 50 e-/Å^2 Gain reference (gain_ref.mrc) is included. Picked particle coordinates are provided in run_data.star.

data/temp6_tif-frames_fixed/*.tif data/temp6_tif-frames_fixed/run_data.star